Compound for treating oxidative stress in canines

Abstract

Managing and treating elevated OS biomarkers in mammals such as companion animals with at least one of the supplements alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E, or selenium. Other compounds are Zinc, Vitamin E, Vitamin C, Quercetin, L-glutamine and Robuvit. Diagnosing an oxidative stress (OS) in a mammal detects an OS biomarker, selectively isoprostane and other antioxidant biomarkers such as HODE, microRNAs, TAC, GSH, MDA, and TNF-alpha. The sample can be saliva.

Claims

1. A method of treating a dog with a diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker comprising: a. mixing ingredients of a composition together to form a combination for ingestion by a dog, b. treating the dog with a diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker to achieve a treatment of the dog with the combination composition to reduce an isoprostane marker indicative of oxidative stress (OS) in the dog with a diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker, the composition being a combination of at least vitamin D, and selenium ingredients, to lower the level below 1.75 ng/ml, and c. reducing the isoprostane biomarker in the dog so treated by the composition combination after five to six months to a level below 1.75 ng/ml, wherein the isoprostane level is reduced by at least 40%, the reduction being determined by i. measuring the biomarker level in saliva from the dog with the previously diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker.

2. The method for treating oxidative stress (OS) as claimed in claim 1 including treating the dog with a diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker with the combination of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, the combination thereby consisting of vitamin D, selenium, alpha-lipoic acid, co-enzyme Q-10, green tea, and turmeric.

3. The method for treating oxidative stress (OS) as claimed in claim 1 including treating an elevated HODE OS biomarker in the dog.

4. The method for treating oxidative stress (OS) as claimed in claim 1 including mixing in the combination alpha-lipoic acid, co-enzyme Q-10, green tea, and turmeric, the combination thereby consisting of the vitamin D, the selenium, alpha-lipoic acid, co-enzyme Q-10, green tea, and turmeric.

5. The method for treating oxidative stress (OS) as claimed in claim 1 including creating in a dosage of about 50 g the combination consisting of Vitamin D3 in an amount of about 75 mcg; Selenium in an amount of about 10 mcg; Green Tea in an amount of about 100 mg; Curcumin-20% total curcuminoids in an amount of about 100 mg; Alpha Lipoic Acid in an amount of about 20 mg and CoQ10 in an amount of about 10 mg.

6. A method of treating a dog with a diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker comprising: a. mixing ingredients together in a combination for ingestion by a dog, and b. treating the dog with a diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker with the combination composition to reduce an isoprostane marker indicative of oxidative stress (OS) in the dog with a diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker, the composition being a combination consisting of vitamin D, selenium, alpha-lipoic acid, co-enzyme Q-10, green tea, and turmeric ingredients, to lower the level below 1.75 ng/ml in the dog with the previously diagnosed level above 1.75 ng/ml of the OS isoprostane biomarker.

7. The method for treating oxidative stress (OS) as claimed in claim 6 including screening a saliva sample of a dog to detect a level of at least an OS isoprostane biomarker.

8. The method for treating oxidative stress (OS) as claimed in claim 6 wherein the level is reduced by at least 40%.

9. The method for treating oxidative stress (OS) as claimed in claim 6 including reducing the isoprostane biomarker in the dog so treated by the composition combination after five to six months to a level below 1.75 ng/ml.

Description

DETAILED DESCRIPTION

(1) Different biomarkers are described and different abbreviations below are as follows.

(2) IsoP, isoprostane(s); AOX, antioxidant capacity; CRP, C-reactive protein; GSH, glutathione; MDA, malonaldehyde; NO, nitric oxide; TAC, total antioxidant capacity; transcription factor Nrf-2 (Nuclear factor erythroid 2-related factor 2), TNF-alpha, tumor necrosis factor-alpha, and HODE, hydroxyoctadecadienoic acid.

(3) The disclosure provides a straightforward, reliable assay for OS biomarkers and for antioxidant capacity of biological fluids.

(4) The present disclosure relates to a test for levels of oxidized lipid biomarkers, selectively isoprostanes, HODE, microRNA and other biomarkers for animals, particularly companion animals and more particularly dogs, cats or horses using saliva that permits the rapid, accurate, non-invasive quantitative screening for biolipids, selectively in the animal.

(5) The saliva-based test assay quantifies the isoprostane, HODE and microDNA levels in dog saliva to determine if the pet's body is undergoing harmful OS. OS creates reactive oxygen species (ROS) causing cells to undergo damage and release biomarker lipids and enzymes that lead to tissue inflammation, infections, periodontal disease, obesity and even cancers. However, free radicals themselves are so reactive and short-lived that direct measurement is not possible. Thus, isoprostane and HODE levels serve as reliable surrogate biomarkers for the presence of ROS.

(6) The saliva-based tests of this disclosure are novel isoprostane, HODE and microRNA tests, and are examples of a set of unique biomarker tests for pets that can be measured in saliva.

(7) For quantitative testing, an animal's serum or saliva or other bodily fluid is added to the ELISA microtiter plate or other immunoassay platforms such as but not limited to lateral flow, or latex or bead agglutination, which measures the presence of cellular oxidative stress.

(8) Once collected at or received by the lab, the blood serum or saliva or other bodily fluid sample is then screened using the ELISA method or other immunoassay platforms such as but not limited to lateral flow, or latex or bead agglutination, which measures the presence of OS.

(9) Forms of biological fluid, other than saliva, for instance urine, tears, sweat, or milk or other mucosal secretions can be used.

(10) The detection of cellular oxidative stress can be performed with an immunoassay. Immunoassays include, but are not limited to, ELISA test, RIA test, latex agglutination, beads assay, and proteomic assays. A preferable immunoassay is the ELISA test. Other immunoassays can be used and the choice of immunoassay can be determined by one of ordinary skill in the art.

(11) A method for diagnosing OS in a mammal such as a human, dog or other companion animal comprises the steps of collecting a sample of saliva; screening the sample to detect a level of at least one of a number of OS markers and detecting and diagnosing the presence of OS based on the level of one or more markers.

(12) A method for diagnosing OS in a mammal such as human, dog or other companion animal comprises the steps of collecting a sample of saliva; screening the sample to detect a level of at least one isoprostane, and detecting and diagnosing the presence of OS based on the quantitative level of the isoprostane.

(13) A method for diagnosing OS in a mammal such as human, dog or other companion animal comprises the steps of collecting a sample of saliva; screening the sample to detect a level of at least one HODE, and detecting and diagnosing the presence of OS based on the quantitative level of the HODE.

(14) The method further comprises collecting a first testing portion of the saliva sample and wherein the first testing portion is the sample for use in the screening step. [Glucuronidase pretreatment is not required for saliva samples.]

(15) The method includes the screening step utilizes an enzyme-linked immunosorbent assay (ELISA) testing system to detect the level of the saliva-based OS marker.

(16) Results

(17) After completing the initial clinical trial studies, analyzing 282 clinical patient samples; 79 of them (35%) were positive, having isoprostane biolipid levels, for example, above the normal reference range we have established (i.e., 0.5-1.75 ng/mL of saliva).

(18) Of the 79 positive testing dogs, there were: 34% spayed females, 32% neutered males, 21% intact males and 15% intact females. The ages ranged from 4 months to 15 years, although most were middle aged or older. The weight range was 4-143 pounds, with 84% being medium to large or giant in size; no breed type predominated.

(19) The diets fed the 79 positive dogs included: 40% ate only a commercial raw diet; 20% only a commercial dry kibble; 13% a home cooked or home prepared raw diet, and 3% ate a combination of a commercial kibble and raw.

(20) Of the 79 positive dogs, 38 also had saliva-based profiles run for food sensitivity and intolerances to 24 foods. Interestingly, only 3 of these 38 dogs had Saliva based test results that were reactive (20 or more foods). These results suggest that dogs with clinical issues related to intense itching, scratching, chewing, and bowel irritability had relatively few identified foods as the culprits. Environmental exposure to inhalants, fleas, ticks, mites and other insects as well as contact reactants could be involved; and 3 dogs were taking an isooxazoline parasiticide.

(21) Positive testing dogs should be retested in about 6 months after being on foods and supplements designed to lower their OS. The beneficial outcome of 50 isoprostane positive samples revealed reduced isoprostane levels when retested 5-6 months after taking the supplements listed below.

(22) Management and Treatment

(23) Management and treatments measures to alleviate elevated oxidative stress biomarkers of at least isoprostane and/or HODE, and their clinical expression in companion animals such as dogs or cats, and other animal species are described below. Use of supplements for isoprostane and HODE can be used for any other biomarkers of OS such as MicroRNAs PGF2a, MDA, TAC and others. Supplements that are used to bring down high levels of isoprostane and/or HODE are: Alpha-Lipoic acid Carnitine Co-Enzyme Q-10 Ginger Green tea Licorice Milk thistle, and a few more like garlic and honey. Resveratrol (as a natural supplement or as food like blueberries and cranberries) Selenium Soybeans Tomatoes Turmeric (curcumin)—without black pepper for pets Vitamin D Vitamin E

(24) A first compound is a veterinary nutritional compound of at least vitamin D and selenium. This first supplement can include alpha-lipoic acid, co-enzyme Q-10, green tea, and turmeric. The first compound can be for treating an elevated isoprostane OS biomarker level in a dog with supplements. The isoprostane biomarker level in the dog so treated is reduced by at least 40% after five to six months to a level below 1.75 ng/ml as measured in saliva from the dog. This first supplement can include Vitamin D3, Selenium, alpha-lipoic acid, co-enzyme Q-10, green tea, and turmeric.

(25) One formulation of a first compound for a maintenance dosage of 0.50 g Vitamin D3 75 mcg Selenium 10 mcg Green Tea 100 mg Curcumin-20% total curcuminoids 100 mg Alpha Lipoic Acid 20 mg CoQ10 10 mg

(26) A second compound is a veterinary nutritional compound of at least L-glutamine and Robuvit (trademark) namely French oak extract. This first supplement can include zinc, vitamin E, vitamin C and quercetin.

(27) The second compound can be for treating an elevated OS biomarker level in a dog with supplements. The biomarker level in the dog so treated is reduced selectively by at least 40% after about five to six months from an elevated level, for instance an isopostrane biomarker level can be reduced to a level below 1.75 ng/ml as measured in saliva from the dog. This second supplement can include L-glutamine, Robuvit (trademark) namely French oak extract, zinc, vitamin E, vitamin C and quercetin.

(28) A second compound is a veterinary nutritional compound of: Zinc Vitamin E Vitamin C Quercetin L-glutamine Robuvit (French oak extract).

(29) A formulation for a maintenance dosage of the second compound is Zinc 50 mg Vitamin E 67 mg Vitamin C 500 mg Quercetin 250 mg L-glutamine 500 mg Robuvit (French oak extract) 100 mg

(30) The amount of each one of these first and second compounds may change up or down by about 25% depending, for instance, on the size of the dog: The larger the dog the greater some of the elements of the compound, and conversely the smaller the dog the less some of the elements of the compounds.

(31) A synergistic biological effect of mutual benefit, based upon the principle of cooperative symbiosis, is achieved by this group of ingredients.

(32) Two or more of Vitamin D3, Selenium, Alpha-Lipoic acid; Carnitine, Co-Enzyme Q-10, Curcumin, and Green tea produce enhanced affects and thus are synergistic.

(33) Two or more of Zinc, Vitamin E, Vitamin C, Quercetin, L-glutamine, and Robuvit (French oak extract) produce enhanced affects and thus are synergistic.

(34) The synergy results from the interaction between nutrients, their absorption and bioavailability in the body which in this situation is positive and provides health benefits. Evidence-based studies have illustrated that the active components in the nutritional supplements of the disclosed supplements work together synergistically to enhance their functional properties in preventing diseases and betterment of health. The role of the disclosed nutritional supplements working together enhances treatment and prevention of chronic diseases, such as cardiovascular and periodontal disease, infections, dysbiosis, diabetes, obesity, and cancers. This combination synergic effect is a scientific advance in nutritional therapy and significantly reduces high levels of isoprostane and/or HODE more effectively than when the supplements are separately used.

(35) Synergistic effects can be obtained with Garlic and honey, Ginger, Resveratrol (as a natural supplement or as food like blueberries and cranberries); Carnitine, licorice, Milk thistle, Soybeans; and Tomatoes.

(36) Each of these individually produce overtly similar effects, they display greatly enhanced effects when given in combination. The combined effect is greater than that predicted by their individual potencies, and thus the combination is synergistic.

(37) The compound of the disclosure includes treating an elevated isoprostane OS biomarker level with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

(38) The method of the disclosure includes treating an elevated isoprostane OS biomarker level and an elevated HODE OS biomarker level with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

(39) The compound of the disclosure includes treating an elevated HODE OS biomarker with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

(40) The compound of the disclosure includes treating an elevated OS biomarker being at least TAC, GSH, MDA, TNF-alpha, NO, and microRNA with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

(41) The compound of the disclosure includes treating an elevated isoprostane OS biomarker level with at least alpha-lipoic acid, co-enzyme Q-10, green tea, milk thistle, garlic, ginger, licorice, carnitine, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, or selenium.

(42) The compound of the disclosure includes treating an elevated HODE OS biomarker with at least alpha-lipoic acid, co-enzyme Q-10, green tea, milk thistle, garlic, ginger, licorice, carnitine, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, or selenium.

(43) The compound of the disclosure includes treating an elevated isoprostane OS biomarker level with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

(44) The compound of the disclosure includes treating an elevated isoprostane OS biomarker level and an elevated HODE OS biomarker level with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of: alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

(45) The compound of the disclosure includes treating an elevated HODE OS biomarker level with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

(46) The compound of the disclosure includes treating an elevated OS biomarker level being at least TAC, GSH, MDA, TNF-alpha, NO, and microRNA with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

(47) The compound of the disclosure includes treating an elevated OS biomarker level being at least TAC, GSH, MDA, TNF-alpha, NO, and microRNA with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of: alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

(48) The compound of the disclosure includes treating an elevated OS biomarker level being at least TAC, GSH, MDA, TNF-alpha, NO, and microRNA with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of a veterinary nutritional compound of French oak extract, L-glutamine, Zinc, Vitamin E, Vitamin C, and Quercetin.

(49) The compound of the disclosure includes treating an elevated isoprostane or HODE OS biomarker level with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of a veterinary nutritional compound of French oak extract, L-glutamine, Zinc, Vitamin E, Vitamin C, and Quercetin.

(50) The amount of each one of these compounds may change up or down by about 25% depending, for instance, on the size of the dog: The larger the dog the greater some of the elements of the compound, and conversely the smaller the dog the less some of the elements of the compounds.

(51) The proportional decrease in the biomarker level of the oxidative stress, being the value of the isoprostane and/or HODE has been at least 40% and in some cases as much as 90% after about 5-6 months of ingestion of the compounds of the disclosure.

(52) The compound is formulated with the ingredients being mixed together in powdered form and being for ingestion and digestion by the mammal, selectively an animal at least once or twice per day together with the food that is being eaten. In this format, the powder given separately or mixed into the food prior to the food being given to the mammal such as human or animal.

(53) General

(54) Many different formats are possible. In the specification, there have been disclosed typical embodiments of the disclosure. Although specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limitation, the scope of the disclosure being set out in the claims. It is therefore to be understood that within the scope of the appended claims the disclosure may be practiced otherwise than as described.